Alprazolam (Xanax®) HPLC Method Transferred – AppNote

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June 5, 2014
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AppNotes ANP/HILIC Methods / AppNotes Cogent Diamond Hydride / AppNotes Library / AppNotes Pharmaceuticals / ARTICLES, APPNOTES, FAQ

Benefit of 2.2um Column Analyzing Alprazolam with HPLC

In this AppNote, we show that a transfer from a 4µm to 2.2µm Column, higher efficiency is observed with the smaller particle size Column. This can be very useful for analyses requiring detection at low concentrations.

The Peak shape is very sharp using the 2.2um Column (Figure. A) and is further sharpened with the use of a gradient. Furthermore, the method conditions are LCMS compatible and could be applied to a variety of applications involving Alprazolam analysis such as clinical assays of Plasma extracts.

Peak:
Alprazolam (API)

Method Conditions
Columns:
—Fig. A: Cogent Diamond Hydride 2.o™, 2.2µm, 120Å
—Fig. B: Cogent Diamond Hydride™, 4µm, 100Å
Catalog Nos.:
—Fig. A: 70200-05P-2;
—Fig. B: 70000-05P-2
Dimensions:
—Fig. A: 2.1 x 50 mm
—Fig. B: 4.6 x 50 mm
Mobile Phase:
—A: DI Water / 0.1% Formic Acid (v/v)
—B: Acetonitrile / 0.1% Formic Acid (v/v)
Gradient:

Time (minutes)	%B
0	95
1	95
6	50
7	95

Post time: 3 minutes
Injection vol.: 0.2 µL
Flow rate: 0.29 mL / minutes
Detection: UV @ 254 nm
Sample: 15 mg strength Morphine Sulfate tablet was ground and weighed in a 25 mL Volumetric Flask. A portion of 50 / 50 solvent A / solvent B diluent was added and the flask was sonicated for 10 minutes. It was then diluted to mark and filtered with a 0.45µm Nylon Syringe Filter (MicroSolv Tech Corp.).
t0: 0.6 minutes